Characterization of dual specificity protein kinase from maize seedlings.

Abstract

A protein kinase of 57 kDa, able to phosphorylate tyrosine in synthetic substrates pol(Glu4,Tyr1) and a fragment of Src tyrosine kinase, was isolated and partly purified from maize seedlings (Zea mays). The protein kinase was able to phosphorylate exogenous proteins: enolase, caseins, histones and myelin basic protein. Amino acid analysis of phosphorylated casein and enolase, as well as of phosphorylated endogenous proteins, showed that both Tyr and Ser residues were phosphorylated. Phosphotyrosine was also immunodetected in the 57 kDa protein fraction. In the protein fraction there are present 57 kDa protein kinase and enolase. This co-purification suggests that enolase can be an endogenous substrate of the kinase. The two proteins could be resolved by two-dimensional electrophoresis. Specific inhibitors of typical protein-tyrosine kinases had essentially no effect on the activity of the maize enzyme. Staurosporine, a nonspecific inhibitor of protein kinases, effectively inhibited the 57 kDa protein kinase. Also, poly L-lysine and heparin inhibited tyrosine phosphorylation by 57 kDa maize protein kinase. The substrate and inhibitor specificities of the 57 kDa maize protein kinase phosphorylating tyrosine indicate that it is a novel plant dual-specificity protein kinase.

A comparative proteome and phosphoproteome analysis of differentially regulated proteins during fertilization in the self‐incompatible species Solanum chacoense Bitt.

“…lase has also been shown to be the substrate of a 57 kD dualspecific protein kinase, being mainly phosphorylated on serine and tyrosine residues [55], thus confirming our identification of enolase as a phosphoprotein. The levels of expression of the putative succinate dehydrogenase flavoprotein alpha subunit does not change as judged from total staining, while its phosphorylation state increases as judged with ProQ Diamond staining ( Figs.…”